, Volume 21, Issue 5, pp 578–593 | Cite as

Combinatorial treatment with anacardic acid followed by TRAIL augments induction of apoptosis in TRAIL resistant cancer cells by the regulation of p53, MAPK and NFκβ pathways

  • M. Harsha Raj
  • B. Yashaswini
  • Jochen Rössler
  • Bharathi P. Salimath


TRAIL, an apoptosis inducing cytokine currently in phase II clinical trial, was investigated for its capability to induce apoptosis in six different human tumor cell lines out of which three cell lines showed resistance to TRAIL induced apoptosis. To investigate whether Anacardic acid (A1) an active component of Anacardium occidentale can sensitize the resistant cell lines to TRAIL induced apoptosis, we treated the resistant cells with suboptimal concentration of A1 and showed that it is a potent enhancer of TRAIL induced apoptosis which up-regulates the expression of both DR4 and DR5 receptors, which has been observed in the cellular, protein and mRNA levels. The death receptors upregulation consequent to A1 treatment was corroborated by the activation of p53 as well as phosphorylation of p38 and JNK MAP kinases and concomitant inactivation of NFκβ and ERK signaling cascades. Also, A1 modulated the expression of key apoptotic players like Bax, Bcl-2 and CAD along with the abatement of tumor angiogenesis in vivo in EAT mouse model. Thus, post A1 treatment the TRAIL resistant cells turned into TRAIL sensitive cells. Hence our results demonstrate that A1 can synergize TRAIL induced apoptosis through the upregulation of death receptors and downregulation of anti-apoptotic proteins in cancer context.


Anacardic acid TRAIL Death receptors Apoptosis 



Anacardic acid


TNFα related apoptosis inducing ligand


Death receptors

DR4, DR5

Death receptor 4 and 5

DcR1, DcR2

Decoy receptor 1 and 2


FAS ligand


Tumor necrosis factor α


Bcl-2-associated X protein


B-cell lymphoma 2 protein


Caspase activated DNase



We acknowledge funding by Department of Biotechnology, New Delhi, India, BT/IN/German/06/BPS/2010 and BMBF-IND 10/026, to BPS and JR respectively, University Grants Commission-Government of India No. f 4-1/2013SAP II and f.No. 14/4/2012(NS/PE). The discussions and help given by Prof. Dr. Regine Süss and Dr. V. Rengaswamy for liposome studies is acknowledged.

Supplementary material

10495_2016_1223_MOESM1_ESM.tif (2.7 mb)
Supplementary material 1 (TIFF 2773 kb) Sup Fig. 1 (a) Liposome size was determined by dynamic light scattering using a zeta potential/particle sizer of freshly prepared liposomes was performed by diluting in isotonic HBS buffer. (b) In vitro cytotoxicity of A1-liposome formulation and free drug A1 (100 µg) on 8 human tumor cell lines: 5x104 cells were seeded in 46 well plates and treated with A1-liposome formulation, A1 alone, free liposomes and untreated cells for 12 h. The cytotoxicity was assessed by trypan blue dye exclusion method


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • M. Harsha Raj
    • 1
  • B. Yashaswini
    • 1
  • Jochen Rössler
    • 2
  • Bharathi P. Salimath
    • 1
  1. 1.Molecular Oncology Lab, Department of Studies in BiotechnologyUniversity of MysoreMysoreIndia
  2. 2.Clinic IV: Pediatric Hematology and Oncology, Center of Pediatrics and Adolescent MedicineUniversity Medical HospitalFreiburgGermany

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